Abstract Stimuli associated with drugs have a powerful influence over the behavior. Many studies have established the ability of drug-related cues and contexts to induce relapse after a period of abstinence. Importantly, exposure to drug-associated cues can also produce specific interoceptive effects related to drug craving, such as feelings of increased tension, jitteriness, and changes in heart rate. However few studies have examined how interoceptive responses to these cues can then become drivers of behavior. The current experiments propose to use a highly relevant behavioral model of context-induced reinstatement combined with manipulation of neural circuitry likely to be involved in interoceptive perception using viral chemogenetic technology. In these experiments, rats will be trained to self-administer alcohol in a specific context that will become associated with the drug. Subsequently, rats will undergo extinction in a separate context where responses do not result in delivery of alcohol. Then, rats will be tested in the original alcohol-associated context in a two-phase reinstatement procedure consisting of a seeking phase where lever presses have no programmed consequence (i.e., seeking phase) and then a re-initiation of self-administration phase where responses will again be rewarded (i.e., re-initiation of self-administration phase). This design allows for examination of multiple factors that can contribute to relapse. Using this design, Aim 1 will determine the functional role of the nucleus reuniens (RE) to anterior insula (AI) projection with virus infusions of Gi-coupled hM4Di Designer Receptors Exclusively Activated by Designer Drugs (DREADDs) into the RE followed by implantation of bilateral cannulae targeting the AI. This circuit will be silenced on the reinstatement test day by activating the Gi DREADDs via delivery of the ligand clozapine-N-oxide (CNO) into the AI. I hypothesize that silencing this circuit will disrupt processing of interoceptive cues that contribute to alcohol seeking and will increase responding upon re-initiation of self-administration. Aim 2 will use the same viral strategy to silence the projection from the nucleus of the solitary tract (NTS) to the RE. I hypothesize that Gi DREADD activation in this circuit will both disrupt interoceptive cues that contribute to alcohol seeking behavior and re-initiation of alcohol self-administration. Altogether, the experiments in this application seek to demonstrate that silencing of brain circuits likely to be involved in the processing of peripheral body state information disrupts use of this information as an interoceptive cue that contributes to alcohol use disorder and relapse.